Non-transgenic soft textured tetraploid wheat plants having grain with soft textured endosperm, endosperm therefrom and uses thereof

ABSTRACT

The present invention relates to non-transgenic tetraploid wheat plants having soft textured endosperm, methods for constructing said non-transgenic plants using cytogenetics and classical breeding techniques, endosperm therefrom and uses thereof.

RELATED APPLICATIONS

This application is claims priority to and is a Continuation of U.S.patent application Ser. No. 12/538,858, filed Aug. 10, 2009, which isincorporated herein by reference in its entirety.

FIELD OF THE INVENTION

The invention relates to wheat breeding, new non-transgenic tetraploidwheat cultivars having grain with soft textured endosperm, methods ofproducing the new non-transgenic tetraploid wheat cultivars and usesthereof.

BACKGROUND OF THE INVENTION

Wheat is one of the most important food crops in the world, and is adominant grain of world commerce. As is well known in the art, thehardness of wheat grain varies between different wheat cultivars. Grainhardness refers to the texture of the kernel (caryopsis), in particularwhether the endosperm is physically hard or soft. Grain texture, or“hardness”, influences processing, including e.g., milling, thecharacteristics of milled granular products of different wheat grainvarieties, and how those varieties and granular products are used infoods.

Different grain textures are exploited to produce a wide variety ofgranular products including e.g., flours, semolinas, farinas, as well asa wide variety of food products. Generally speaking, hard wheat(Triticum aestivum) is used for bread and pasta, whereas soft wheat (T.aestivum) is used for cookies, cakes and pastries (Morris & Rose, CerealGrain Quality, Chapman & Hall, New York, N.Y., pp. 3-54 (1996)).Typically, the very hard durum wheat (T. turgidum ssp. durum) is used inpasta, bread, couscous and bulgur.

Soft wheat typically produces a finer granulation upon milling orgrinding, hard wheat a coarser granulation, and durum wheat a verycoarse granulation. Although granulation is controlled somewhat by theart of milling, it is still highly limited by the inherent hardnesscharacteristics of the wheat variety.

Durum wheat is universally recognized as the ideal raw material for theproduction of quality pasta products. In particular, durum wheat is highin protein and gluten, both of which are necessary components for pastamaking. As noted above, durum wheat typically has a grain with a veryhard textured endosperm. Consequently, it is typically milled into agranular meal of coarse particle size called semolina. Finergranulations can be produced however to produce a durum meal or flour ofa particular granularity, more energy is required for milling than isrequired for the milling of other wheat having grain with softertextured endosperm. Thus, it is particularly difficult to obtain afinely granulated flour from durum wheat without incurring excessivestarch damage and expending considerable energy. Yet, finer granulationsare increasingly preferred by modern pasta manufacturers.

Starch damage influences water absorption and other dough properties,and despite other desirable factors e.g., protein content, pastaprepared with highly damaged starch is typically of inferior qualitye.g., is sticky and has poor “mouth feel”, and is therefore consideredby most people to be unacceptable. See e.g., Pasta and Noodle TechnologyJames E. Kruger et al. eds., American Association of Cereal Chemists(1996).

Thus, the energy expended in milling the durum wheat kernels intosemolina and flour must be carefully controlled so as to achieve thedesired granulation with minimal starch damage. Therefore, to take fulladvantage of the beneficial and desirable characteristics of durumwheat, what is needed in the art is a durum wheat having grain with softtextured endosperm. Indeed, such a wheat cultivar would inter alia,produce soft textured wheat grain which could be ground into variousgranulations including finely textured flour without extensive amountsof starch damage an/or which could be ground more easily into semolinawith much less energy expended.

Transgenic soft kernel wheat is known (see e.g., U.S. Pat. No.6,596,930). However, there are barriers to widespread use of geneticallymodified cultivars. Indeed, genetically modified crops face highregulatory and research costs, and are opposed by many NGOs, and severalgovernments, particularly within the European Community and Japan.

Therefore, a durum wheat having grain with soft textured endosperm thatis also non-transgenic would meet the need for soft textured durum, andwould also get around the barriers to widespread use that facegenetically modified crops.

Fortunately, as will be clear from the following disclosure, the presentinvention provides for these and other needs.

SUMMARY OF THE INVENTION

One exemplary embodiment of the invention provides a non-transgenictetraploid wheat plant having grain with soft textured endosperm whereinthe non-transgenic tetraploid wheat plant having grain with softtextured endosperm has functional Ph1 and diploid like behavior inmeosis. In one exemplary embodiment, the non-transgenic tetraploid wheatplant is a durum wheat plant. In another exemplary embodiment, thenon-transgenic tetraploid wheat plant is capable of serving as a parentin a genetic cross.

Another exemplary embodiment of the invention provides a seed of anon-transgenic tetraploid wheat plant having grain with soft texturedendosperm, wherein the seed produces a tetraploid wheat plant havinggrain with soft textured endosperm. In another exemplary embodiment, theinvention provides a non-transgenic tetraploid wheat plant having grainwith soft textured endosperm, or a part thereof, produced by growing theseed.

Another exemplary embodiment of the invention provides a tissue cultureof regenerable cells produced from a non-transgenic tetraploid wheatplant having grain with soft textured endosperm. In another exemplaryembodiment, the invention provides a protoplast produced from the tissueculture a tissue culture of regenerable cells produced from anon-transgenic tetraploid wheat plant having grain with soft texturedendosperm.

Another exemplary embodiment of the invention provides a non-transgenictetraploid wheat plant having grain with soft textured endosperm, or apart thereof, having all the physiological and morphologicalcharacteristics of the variety Soft Svevo durum wheat WAS 080240001(hereinafter, Soft Svevo), representative seed of such line having beendeposited under ATCC Accession No. PTA-10087. In one exemplaryembodiment, the non-transgenic tetraploid wheat plant is capable ofserving as a parent in a genetic cross. In another exemplary embodiment,a seed of the non-transgenic tetraploid wheat plant produces anon-transgenic tetraploid wheat plant having grain with soft texturedendosperm. In another exemplary embodiments the non-transgenictetraploid wheat plant is used to produce a tissue culture ofregenerable cells. In another exemplary embodiment a protoplast isproduced from the tissue culture of regenerable cells. In still anotherexemplary embodiment, the invention provides a non-transgenic tetraploidwheat plant having grain with soft textured endosperm, regenerated fromthe tissue culture of regenerable cells produced from a non-transgenictetraploid wheat plant having grain with soft textured endosperm.

Another exemplary embodiment of the invention provides a hybrid wheatplant, wherein the lineage of at least one parent plant comprises anon-transgenic tetraploid wheat plant having grain with soft texturedendosperm, having all the physiological and morphologicalcharacteristics of the variety Soft Svevo, representative seed of suchline having been deposited under ATCC Accession No. PTA-10087. In oneexemplary embodiment the at least one parent plant of the hybrid wheatplant is the non-transgenic tetraploid wheat plant having grain withsoft textured endosperm, with all the physiological and morphologicalcharacteristics of the variety Soft Svevo, representative seed of suchline having been deposited under ATCC Accession No. PTA-10087.

Another exemplary embodiment of the invention provides a non-transgenictetraploid wheat plant having grain with soft textured endosperm havingATCC Accession No. PTA-10087 or a selfed progeny thereof or an F1 hybridthereof wherein the non-transgenic tetraploid wheat plant has grain withsoft textured endosperm.

Another exemplary embodiment of the invention provides a semolina flourcomprising soft textured endosperm from a non-transgenic tetraploidwheat plant having grain with soft textured endosperm. In one exemplaryembodiment, the semolina flour of is made from a non-transgenictetraploid wheat plant having ATCC Accession No. PTA-10087 or a progenythereof.

Other features, objects and advantages of the invention will be apparentfrom the detailed description which follows.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Miag Multomat flour mill schematic flow diagram.

FIG. 2. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the Italian durum wheat cultivar Svevo (filledtriangles) with non-transgenic Langdon durum homoeologous translocationline ‘674’ F3:4 plants as male. Lines indicate suggested delineations inkernel texture phenotypic classes. The circled symbol identifies softdurum progeny translocation line ‘152’ which was selected and used todevelop ‘Soft Svevo Durum Wheat WAS 080240001’ which was deposited withthe American Type Culture Collection under Accession number PTA-10087.

FIG. 3. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the Italian durum wheat cultivar Creso (filledcircles) with non-transgenic Langdon durum homoeologous translocationline ‘674’ F3:4 plants as male. Lines indicate suggested delineations inkernel texture phenotypic classes.

FIG. 4. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the Italian durum wheat cultivar Svevo (filledtriangles) with non-transgenic Langdon durum homoeologous translocationline ‘675’ F3:4 plants as male. Lines indicate suggested delineations inkernel texture phenotypic classes.

FIG. 5. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the Italian durum wheat cultivar Creso (filledcircles) with non-transgenic Langdon durum homoeologous translocationline ‘675’ F3:4 plants as male. Lines indicate suggested delineations inkernel texture phenotypic classes.

FIG. 6. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the Italian durum wheat cultivar Svevo (filledtriangles) with non-transgenic Langdon durum homoeologous translocationline ‘678’ F3:4 plants as male. Lines indicate suggested delineations inkernel texture phenotypic classes.

FIG. 7. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the Italian durum wheat cultivar Creso (filledcircles) with non-transgenic Langdon durum homoeologous translocationline ‘678’ F3:4 plants as male. Lines indicate suggested delineations inkernel texture phenotypic classes.

FIG. 8. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the Italian durum wheat cultivar Svevo (filledtriangles) with non-transgenic Langdon durum homoeologous translocationline ‘679’ F3:4 plants as male. Lines indicate suggested delineations inkernel texture phenotypic classes.

FIG. 9. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the Italian durum wheat cultivar Creso (filledcircles) with non-transgenic Langdon durum homoeologous translocationline ‘679’ F3:4 plants as male. Lines indicate suggested delineations inkernel texture phenotypic classes.

FIG. 10. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the Italian durum wheat cultivar Svevo (filledtriangles) with non-transgenic Langdon durum homoeologous translocationline ‘685’ F3:4 plants as male. Lines indicate suggested delineations inkernel texture phenotypic classes.

FIG. 11. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the Italian durum wheat cultivar Creso (filledcircles) with non-transgenic Langdon durum homoeologous translocationline ‘685’ F3:4 plants as male. Lines indicate suggested delineations inkernel texture phenotypic classes.

FIG. 12. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the Italian durum wheat cultivar Svevo (filledtriangles) with non-transgenic Langdon durum homoeologous translocationline ‘688’ F3:4 plants as male. Lines indicate suggested delineations inkernel texture phenotypic classes.

FIG. 13. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the Italian durum wheat cultivar Creso (filledcircles) with non-transgenic Langdon durum homoeologous translocationline ‘688’ F3:4 plants as male. Lines indicate suggested delineations inkernel texture phenotypic classes.

FIG. 14. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F5 kernels harvested from individual F4 plantsderived by self pollinating three different non-transgenic Langdon durumhomoeologous translocation lines crossed to the Italian durum wheatcultivar Creso (filled circles) and Svevo (filled triangles). Creso(filled square) and Svevo (filled diamond) parents are shown. Linesindicate suggested delineations in kernel texture phenotype classes.

FIG. 15. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of BC3F2 kernels harvested from individual BC3F1plants derived from Svevo/674 line no. 152 (filled circles and thefilled square). The selected progeny (filled square is Soft Svevo durumwheat WAS 080240001, representative seed of such line having beendeposited under ATCC Accession No. PTA-10087) is shown as are two Svevo(filled triangle) parent plants.

FIG. 16. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the North American durum wheat cultivar Kyle withnon-transgenic Langdon durum homoeologous translocation line 678 plantsas male (filled circles). Kyle (filled square) parent plant is shown.The line indicates a suggested delineation in kernel texture phenotypeclasses. The circled progeny line was used in segregation analysis shownin FIG. 18. The cross symbol identifies the five lines selected forback-crossing to Kyle (two from 678 cross, three from 683 cross).

FIG. 17. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the North American durum wheat cultivar Kyle withnon-transgenic Langdon durum homoeologous translocation line 683 plantsas male (filled triangles). Kyle (filled square) parent plant is shown.The line indicates a suggested delineation in kernel texture phenotypeclasses. The circled progeny line was used in segregation analysis shownin FIG. 18. The cross symbol identifies the five lines selected forback-crossing to Kyle (two from 678 cross, three from 683 cross).

FIG. 18. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F3 kernels harvested from individual F2 plantsderived from a single F1 plant, and produced by crossing the NorthAmerican durum wheat cultivar Kyle with non-transgenic Langdon durumhomoeologous translocation line 683. The lines indicate suggesteddelineations in kernel texture phenotype classes.

FIG. 19. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of BC1F2 kernels harvested from individual BC1F1plants derived by crossing the North American durum wheat cultivar Kylewith non-transgenic Langdon durum homoeologous translocation line 678(filled circles) and 683 (filled triangles). Kyle (filled square) parentplant is shown. The lines indicate suggested delineations in kerneltexture phenotype classes.

FIG. 20. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the North American durum wheat cultivar Havasu withnon-transgenic Soft Svevo durum wheat WAS 080240001 (filled circles).Havasu (filled triangles) parent plants are shown, and a soft hexaploidwheat control (filled square).

FIG. 21. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F3 kernels harvested from individual F2 plantsderived by crossing the North American durum wheat cultivar Havasu withnon-transgenic Soft Svevo durum wheat WAS 080240001 (filled circles).Havasu (filled triangles) parent plants are shown, as is a soft (filledsquare) and hard (filled diamond) hexaploid wheat controls. The linesindicate suggested delineations in kernel texture phenotype classes. BC1seeds were selected from those plants that are circled for advancing.

FIG. 22. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of BC1F3 kernels harvested from individual BC1F2plants derived by crossing the North American durum wheat cultivarHavasu with non-transgenic Soft Svevo durum wheat WAS 080240001 (filledcircles). Havasu (filled triangles) parent plants are shown, as is ahard hexaploid wheat control (filled square).

FIG. 23. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of BC2F3 kernels harvested from individual BC2F2plants derived by crossing the North American durum wheat cultivarHavasu with non-transgenic Soft Svevo durum wheat WAS 080240001 (filledcircles). Havasu (filled triangles) parent plants are shown. BC3F1 seedswere selected from the plant that is circled for advancing.

FIG. 24. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of BC3F2 kernels harvested from individual BC3F1plants derived by crossing the North American durum wheat cultivarHavasu with non-transgenic Soft Svevo durum wheat WAS 080240001. Thelines indicate suggested delineations in kernel texture phenotypeclasses.

FIG. 25. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the North American durum wheat experimental breedingline CA801-721 with non-transgenic Soft Svevo durum wheat WAS 080240001(filled circles). CA801-721 (filled triangle) parent plant is shown. F2seeds were selected from the plant that is circled for advancing.

FIG. 26. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the North American durum wheat experimental breedingline CA801-721 with non-transgenic Soft Svevo durum wheat WAS 080240001(filled circles). CA801-721 (filled triangles) parent plants are shown.

FIG. 27. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the North American durum wheat experimental breedingline CA801-721 with non-transgenic Soft Svevo durum wheat WAS 080240001.

FIG. 28. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the North American durum wheat cultivar Alzada withnon-transgenic Soft Svevo durum wheat WAS 080240001. Alzada (filledtriangles) parent plants are shown. The three plants represented by opencircles were selected for advancing.

FIG. 29. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F3 kernels harvested from individual F2 plantsderived by crossing the North American durum wheat cultivar Alzada withnon-transgenic Soft Svevo durum wheat WAS 080240001. Alzada (filledtriangles) parent plants are shown. BC1 seed from the two plantsrepresented by open circles were selected for advancing.

FIG. 30. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of BC1F2 kernels harvested from individual BC1F1plants derived by crossing the North American durum wheat cultivarAlzada with non-transgenic Soft Svevo durum wheat WAS 080240001. Alzada(filled triangles) parent plants are shown.

FIG. 31. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of BC2F2 kernels harvested from individual BC2F1plants derived by crossing the North American durum wheat cultivarAlzada with non-transgenic Soft Svevo durum wheat WAS 080240001. Alzada(filled triangles) parent plants are shown. BC3F1 kernels from the fourplants with SKCS hardness indexes less than 45 were selected foradvancing.

FIG. 32. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of F2 kernels harvested from individual F1 plantsderived by crossing the North American durum wheat cultivar Strongfieldwith non-transgenic Soft Svevo durum wheat WAS 080240001. Strongfield(filled triangles) parent plants are shown.

FIG. 33. Plot of SKCS kernel texture Hardness Index (HI) versus SKCS HIstandard deviation of BC1F2 kernels harvested from individual BC1F1plants derived by crossing the North American durum wheat cultivarStrongfield with non-transgenic Soft Svevo durum wheat WAS 080240001.Strongfield (filled triangles) parent plants are shown.

DETAILED DESCRIPTION OF THE INVENTION Definitions

The term “plant” as used herein refers to whole plants, plant bodies,plant organs (e.g., leaves, stems, flowers, roots, etc.), seeds, planttissues, plant cells and progeny of same. In an exemplary embodiment, aplant cell includes callus. In another exemplary embodiment, a plantorgan includes a root, a leaf, a flower and/or the like. The term“plant” refers to plants of any variety of ploidy levels, includingpolyploid, diploid, haploid and hemizygous.

The term “transgenic plant” as used herein refers to a plant comprisinga heterologous nucleic acid sequence that codes for or otherwiseinfluences expression of the desirable trait of soft kernel texturewherein the heterologous nucleic acid sequence was introduced into theplant, at some point in its lineage, by genetic engineering techniques.Thus, the term “transgenic plant” refers to tetraploid wheat plantswhich are the direct result of transformation with a heterologousnucleic acid or transgene, and the progeny and decendants of transformedplants which comprise the introduced heterologous nucleic acid ortransgene. In an exemplary embodiment, a “transgenic plant” is atetraploid wheat plant having grain with soft kernel texture wherein thesoft kernel texture is a result of expression of a heterologoustransgene e.g., puroindoline a and/or puroindoline b, that codes forand/or influences the expression of soft kernel texture.

A “non-transgenic tetraploid wheat plant having grain with soft kerneltexture” or a “non-transgenic durum wheat plant having grain with softkernel texture” as used herein, refers to a tetraploid wheat plant thatdoes not comprise heterologous nucleic acid sequences for the desirabletrait of soft kernel texture, either as a direct result oftransformation with a heterologous nucleic acid or transgene, or as aresult of inheritance as the progeny and decendants of transformedplants which comprise the introduced heterologous nucleic acid ortransgene. Typically, the desirable traits of a “non-transgenictetraploid wheat plant having grain with soft kernel texture” e.g., softkernel texture, are produced and transmitted from one generation to thenext by virtue of classical selective breeding techniques without theneed for any genetic engineering techniques. Thus, a “non-transgenictetraploid wheat plant having grain with soft kernel texture” is a wheatplant that is non-transgenic for genes and/or at loci that code for orotherwise influence the desirable soft kernel texture trait. Thus, in anexemplary embodiment, a non-transgenic tetraploid wheat plant havinggrain with soft kernel texture is non-transgenic at the puroindoline aand puroindoline b loci. However, in some exemplary embodiments, anon-transgenic tetraploid wheat plant having grain with soft kerneltexture is transgenic at loci that do not directly code for and/orinfluence the desirable soft kernel trait. Thus, in one exemplaryembodiment, a non-transgenic tetraploid wheat plant having grain withsoft kernel texture is non-transgenic at the puroindoline a andpuroindoline b loci, but is transgenic for another trait, e.g.,herbicide resistance.

The expression “heterologous nucleic acid sequence” or “heterologousgene” as used herein, refers to a gene that is not in its naturalenvironment (in other words, has been altered by the hand of man). In anexemplary embodiment, a heterologous gene is a gene from one speciesthat is introduced into another species. In another exemplaryembodiment, a “heterologous gene” or a “heterologous nucleic acidsequence” is a nucleic acid sequence joined to a regulatory element(s)e.g., a promoter, that is not found naturally associated with the“heterologous gene” or “heterologous nucleic acid sequence”.

The term “tetraploid wheat plant” as used herein, refers to a wheatplant having two sets of paired chromosomes, or four times the haploidnumber of individual chromosomes. The haploid number of wheatchromosomes is seven. Thus, a diploid wheat plant has seven pairs ofchromosomes or 14 chromosomes in total. A “tetraploid wheat plant” hastwo sets of seven pairs, or 28 chromosomes total. Similarly, a“hexaploid wheat plant” has six pairs of seven chromosomes, or 42chromosomes total. In an exemplary embodiment, a tetraploid wheat plantis a durum wheat plant.

The expression “having functional Ph1 and diploid like behavior inmeiosis” as used herein, refers to a wheat plant that exhibits activityof Ph1 (Pairing homoeologous 1) locus as evidenced by the suppression ofpairing among and between homoeologous chromosomes in polyploid wheat,while allowing the normal pairing of homologous chromosomes. Because ofPh1 activity, polyploid wheats behave as functional diploids in theircontrolled and orderly pairing at meiosis. Diploid-like behavior inmeiosis refers to the stable transmission of genetic traits in typicalMendelian genetic ratios, e.g., 1:2:1 segregation of a single gene traitin the F1 generation of a cross between two diploid parents.

The term “soft textured endosperm” “soft textured grain”, “grain withsoft textured endosperm” “soft grain” “soft kernel” “soft kerneltexture” or any gramatically equivalent expression as used herein,refers to grain kernels from a non-transgenic tetraploid wheat plantthat have a hardness index of between about 10 to about 45 as measuredby the Perten SKCS 4100 (Perten Instruments, Reno, Nev. See e.g.,Instruction Manual, Single-Kernel Characterization System, Model SKCS4100; Perten Instruments, Inc.: 6444 South Sixth Street Road,Springfield, Ill. 62707) or by equivalent technology. Kernel hardness istypically expressed as an index/value of −20 to 120.

The kernel hardness associated with grain of hard wheats e.g., harddurum wheats, typically have SKCS values from about 60 to about 100.However, in some exemplary embodiments, the kernel hardness associatedwith grain of hard wheats have SKCS values in a range that is from about50 to about 110. In other exemplary embodiments, the kernel hardnessassociated with grain of hard wheats have an SKCS hardness value in arange that is from about 70 to about 85.

In contrast, the kernel hardness associated with grain of soft wheatse.g., soft durum and soft T. aestivum typically have SKCS hardnessvalues in a range that is from about 10 to about 45. However, in someexemplary embodiments, the kernel hardness associated with grain of softwheats have SKCS hardness values in a range that is from about 0 toabout 55. In other exemplary embodiments, the kernel hardness associatedwith grain of soft wheats have SKCS hardness values in a range that isfrom about 15 to about 35. Thus, in an exemplary embodiment, the grainof a non-transgenic tetraploid wheat plant having soft texturedendosperm has a hardness value that is in a range that is between about10 to about 45. In another exemplary embodiment, the grain of anon-transgenic tetraploid wheat plant having soft textured endosperm hasa hardness value that is in a range that is between about 15 to about 35as measured by the Perten SKCS 4100. And, in still another exemplaryembodiment, the grain of a non-transgenic tetraploid wheat plant havingsoft textured endosperm has a hardness value that is in a range that isbetween about 0 to about 55 as measured by the Perten SKCS 4100.

The term “starch damage” as used herein, refers to mechanical damagethat afflicts starch granules as a result of milling. Typically, starchdamage is manifest as cracks and breaks in the starch granule, and canbe quantitated by any method known in the art (see below).

The term “cross” or “crossing” as used herein refers to a simple X by Ycross, or the process of backcrossing, depending on the context.

The term “backcross” as used herein refers to a process in which abreeder crosses a hybrid progeny line back to one of the parentalgenotypes one or more times.

The terms “disomic”, “ditelosomic”, “heterozygote”, “substitution line”,“filial”, “homoeologous”, “bivalent”, “monosomic”, “monovalent”, “ringbivalent”, “test cross”, “translocation”, “trivalent”, “doubleditelosomic”, “telosomic”, “telomeric”, “nullisomic”, “tetrasomic” orthe plural or adjective form of any of these as used herein havemeanings typically ascribed in the art (see e.g., Rieger R., MichaelisA., Green M. M. (1976) Glosary of Genetics and Cytogenetics. Classicaland Molecular, 4^(th) ed., Springer-Verlag, Berlin; Gill B. S. (1986) Aproposal for wheat chromosome band nomenclature. In: North AmericanWheat Genetic Mapping and Cytogenetic Stocks Workshop, Apr. 17-19, 1986,University of Missouri, Columbia. Ed: Qualset C O and McGuire P E. TheNational Association of Wheat Growers Foundation, Washington, D.C.11-15; Gill B. S., et al. (1991) Genome 34:830-839; Kimber G and SearsER (1968) Nomenclature for the description of aneuploids in theTriticinae. In: Proc 3rd Int Wheat Genet Symp, Ed: Findlay K W andShepherd K W. Canberra, Australia. 468-473; and Raupp W. J., Friebe B.,Gill B. S. Suggested guidelines for the nomenclature and abbreviation ofthe genetic stocks of wheat and its relatives,http://wheat.pw.usda.gov/ggpages/nomenclature.html.

Pedigrees follow the convention of Purdy L. H., Loegering W. Q., KonzakC. F. et al. (1968) Crop Science 8:405-406.

The abbreviations “F1”, “BC1” and higher numerical values of each aloneand in combination (e.g. “BC1F1”) represent “first filial generation”,“first back cross generation”, respectively, etc.

The term “durum flour” as used herein has meaning commonly used in theart and as set forth in CFR §137.220. The expression “Whole durum flour”as used herein has meaning commonly used in the art and as set forth inCFR §137.225 The term “Semolina” as used herein has meaning commonlyused in the art and as set forth in CFR §137.320, and furtherrestrictions relative to the definitions as regards measuringgranulation, ash content, etc. are as provided by current statute.

Taxonomic classifications follow those of M. W. van Slageren, Wildwheats: a monograph of Aegilops L. and Amblyopurum (Jaub. & Spach) Eig(Poaceae) (1994), Agricultural University, Wageningen, the Netherlands.

Milling terms are well-known to those of skill in the art and can befound e.g., in: Posner E. S., Hibbs, A. N. (1997) Wheat Flour MillingAmerican Association of Cereal Chemists, St. Paul, Minn.

I. Introduction

In an exemplary embodiment, the invention provides a non-transgenictetraploid wheat plant having grain with soft kernel texture(soft-textured endosperm). In another exemplary embodiment, theinvention provides non-transgenic tetraploid wheat plants having grainwith soft-textured endosperm, which have a functional Ph1 and diploidlike behavior in meiosis and which therefore are able to serve as aparent in a genetic cross. In still other exemplary embodiments theinvention provides milled products from the disclosed non-transgenictetraploid wheat plants having grain with soft kernel texture. Thus, inaddition to providing a non-transgenic tetraploid wheat plant havinggrain with soft kernel texture the invention provides non-transgenictetraploid wheat plants that are useful as foundation stock for theproduction of new wheat varieties, and provided milled products derivedfrom the soft textured endospem produced by the non-transgenictetraploid wheat plants having grain with soft kernel texture.

Variations in the characteristics and quality of wheat grain influencehow successfully wheat and flour and other granular products perform inconsumer products. Enhancing wheat quality improves processingefficiencies, makes more desirable and more diverse consumer productsand thus ensures the competitiveness of farmers, grain merchandisers,millers, and end processors. The efficient and innovative use of wheatgrain depends on both controlling and exploiting variation in its basicquality traits.

One basic quality trait of importance is kernel texture. Indeed,different wheat grain textures are exploited to produce the wide varietyof granular products e.g., flours, semolinas, farinas, etc. derived fromground wheat. Typically, hard wheat (Triticum aestivum) is used forbread and pasta, soft wheat (T. aestivum) is used for cookies, cakes andpastries (Morris & Rose, Cereal Grain Quality, Chapman & Hall, New York,N.Y., pp. 3-54 (1996)) and very hard durum wheat (T. turgidum ssp.durum) is used e.g., in pasta, bread, couscous and bulgur.

Soft wheat produces a finer granulation upon milling or grinding, hardwheat a coarser granulation, and durum wheat a very coarse granulationreferred to in the art as semolina. As is well known in the art, thoughgranulation is controlled somewhat by milling, it is highly limited bythe inherent hardness characteristics of the wheat variety.

Although it is difficult to obtain a finely granulated durum wheat flourwithout incurring excessive starch damage and expending considerableenergy, finer granulations of durum wheat flour are increasinglypreferred by modern pasta manufacturers. Thus, what is needed in theart, now more than ever, is a durum wheat having grain with soft-kerneltexture.

Soft textured durum wheats are known. However, the available softtextured durum wheats are either transgenic (see e.g., U.S. Pat. No.6,596,930) or are low producing and/or genetically unstable (see e.g.,Liu, C. Y. (1995) Journal of Cereal Science 21:209-213). Marketacceptance of transgenic plants, the produce from transgenic plants, andthe products made from transgenic plants is typically low. Geneticallyunstable wheat plants that are low yielding and which cannot be used forthe development of new wheat varieties are of limited use. Thus, theneed remains for a non-transgenic durum wheat having grain withsoft-kernel texture which has functional Ph1 and diploid like behaviorin meiosis and which therefore can be used as a parent in a geneticcross e.g., as a foundation parent in the development of newnon-transgenic tetraploid wheat plants soft textured endosperm.

Fortunately, the present inventors herein provide such a non-transgenictetraploid wheat plant having grain with soft-kernel texture. Thenon-transgenic tertraploid wheat plants have functional Ph1 and diploidlike behavior in meiosis and are therefore suitable to serve as a parentin a genetic cross. Thus, the non-transgenic tetraploid wheat planthaving grain with soft-kernel texture and having functional Ph1 anddiploid like behavior in meiosis is suitable as foundation stock for thedevelopment of new wheat varieties having grain with soft kerneltexture. In some exemplary embodiments, the non-transgenic tetraploidwheat plants having functional Ph1 and diploid like behavior in meiosishave less than a whole alien chromosome substitution. In other exemplaryembodiments, the non-transgenic tetraploid wheat plants havingfunctional Ph1 and diploid like behavior in meiosis have less than awhole-arm alien chromosome translocation

In an exemplary embodiment, grain from the non-transgenic tetraploidwheat plant having grain with soft-kernel texture is milled to producehigh yields of durum flour with low levels of damaged starch. In anotherexemplary embodiment, grain from the non-transgenic tetraploid wheatplant having grain with soft-kernel texture is milled to produce durumsemolina with low work expenditure compared to semolina prepared fromnormal very hard durum wheat.

II. Genetic Crosses to Produce a Non-Transgenic Soft Durum Wheat Plant.A. General Methods

Methods disclosed herein utilize routine techniques in the field ofwheat genetics, cytogenetics, and milling. Basic terminology in thefield of genetics and cytogenetics can be found e.g., In: Robert C.King, William D. Stansfield, A Dictionary of Genetics, sixth edition2002, Oxford University Press; basic texts in wheat genetics include,e.g., Alain P. Bonjean and William J. Angus, The World Wheat Book AHistory of Wheat Breeding, 2001, Lavoisier Publishing, Paris; K. S.Quisenberry and L. P. Reitz, Wheat and Wheat Improvement, 1967, AmericanSociety of Agronomy, Inc., Madison, Wis.

Techniques and methods for the cytological manipulation and analysis ofwheat are well known to of skill in the art see e.g., Sybenga J. (1992)Cytogenetics in Plant Breeding, Monographs on Theoretical and AppliedGenetics, vol. 17, Springer-Verlag, Berlin; Sharma A. K., Sharma A.(1980) Chromosome Techniques. Theory and Practice, 3^(rd) edn.,Butterworths, London; Morris R., Sears E. R. (1967) The Cytogenetics ofWheat and Its Relatives, pp. 19-87, in Wheat and Wheat Improvement,editors Quisenberry K. S., Reitz L. P., American Society of Agronomy,Madison, Wis.; Gill B. S. (1987) Section 5E: Chromosome Banding Methods,Standard Chromosome Band Nomenclature, and Applications in CytogeneticAnalysis, pp. 243-254, in Wheat and Wheat Improvement, 2^(nd) edn.,editor Heyne E. G., American Society of Agronomy, Madison, Wis.; JoppaL. R. (1987) Section 5F: Aneuploid Analysis in Tetraploid Wheat, pp.255-287, in Wheat and Wheat Improvement, 2^(nd) edn., editor Heyne E.G., American Society of Agronomy, Madison, Wis.; Knott D. R. (1987)Section 7E: Transferring Alien Genes to Wheat, pp. 462-471, in Wheat andWheat Improvement, 2^(nd) edn., editor Heyne E. G., American Society ofAgronomy, Madison, Wis.

Basic terminology and technology in the field of milling can be founde.g., In: Posner E. S., Hibbs, A. N. (1997) Wheat Flour Milling AmericanAssociation of Cereal Chemists, St. Paul, Minn.

B. Genetics of Wheat Having Soft Textured Endosperm

In T. aestivum wheat (bread wheat), differences in grain texture resultfrom the expression of one major gene locus, designated Hardness (Ha)(see e.g., Symes K. J. (1965) Aust J Agric Res. 16:113-123; and Baker R.J. (1977) Crop Sci. 17:960-962). The hardness locus, which is located onthe short arm of chromosome 5D (i.e., 5DS) (see e.g., Mattern, P. J. etal. Proceedings of the 4^(th) International Wheat Genetics Symposium.Columbia, Mo.: Univ. Missouri; 1973. pp. 703-707; and Law, C. N. et al.Seed Protein Improvement by Nuclear Techniques. Vienna, Austria:International Atomic Energy Agency; 1978. pp. 483-502), has two alleles:Ha (soft) and ha (hard). It is generally well accepted that two genespresent at this locus, puroindoline a and puroindoline b, are thecausative agents for grain softness. Mutations in either puroindolinegene result in a partial loss of kernel softness (Morris 2002, Morrisand Bhave 2008), and result in what is known as ‘hard’ T. aestivumwheat.

To have a single gene locus controlling a trait is unusual in polyploidwheat. Indeed, hexaploid wheat (T. aestivum) is an allohexaploid(2n=6x=42 chromosomes; genomes AABBDD). Thus, typically, most genes arepresent in triplicated homoeologous sets, one from each genome. However,in the case of Hardness, the loci on the 5A and 5B chromosomes that werepresent in the diploid progenitors were lost during the formation oftetraploid wheat and thus are not expressed in T. turgidum. Althoughsoftness was restored to hexaploid wheat upon its allohexaploidationwith the diploid Aegilops tauschii (2n=2x=14 chromosomes; genome DD),tetraploid wheats (e.g., durum wheats, T. turgidum L. ssp. durum)(2n=4x=28 chromosomes; genomes AABB), lack the D genome and aretherefore generally much harder textured than soft hexaploid wheat, andgenerally harder than hard hexaploid wheat.

Control of chromosome pairing in wheat during meiosis is under complexgenetic control. One gene locus, Pairing homoeologous (Ph1), whichresides on the long arm of chromosome 5B in T. aestivum and T. turgidum,exerts major control by preventing the pairing of homoeologouschromosomes (e.g., chromosome 5A pairing with 5B or 5D, etc.). Twoinduced mutations in this gene are well known in the art (ph1b in thehexaploid wheat variety Chinese Spring, and ph1c in the tetraploid wheatvariety Cappelli), the presence of either mutation relieves therestriction against homoeologous pairing such that homoeologouschromosomes can pair at some variable frequency and efficiency,resulting in crossing-over and exchange of genetic material (see e.g.,Sears, E. R. (1977) Canadian Journal of Genetics and Cytology19:585-593; Giorgi B. (1978) Mut. Breed. Newsletter 11:4-5; Jauhur P.P., Do{hacek over (g)}ramaci, M., Peterson T. S. (2004) Genome47:1173-1181; Gennaro A., Forte P., Carozza R. et al. (2007) IsraelJournal of Plant Sciences 55:267-276; Qi L., Friebe B., Zhang P., GillB. S. (2007) Chromosome Research 15:3-19; Luo M.-C., Yang Z.-L., Kota R.S., Dvo{hacek over (r)}ák J. (2000) Genetics 154:1301-1308; Jauhar P.P., Riera-Lizarazu O., Dewey W. G. et al. (1991) Theoretical and AppliedGenetics 82:441-449).). This phenomenon is well known in the art as“homoeologous translocation”.

Presence of ph1b affects plant morphology, decreases fertility andreduces male transmission; homozygotes frequently exhibit monosomes andtrisomes, and telocentric chromosomes sometimes occur because of theirregularities associated with homoeologous pairing. Reciprocaltranslocations also occur but are difficult to identify due to formationof multivalents. ph1b varieties are more conveniently maintained as F1heterozygotes, as a single functional copy of a functional Ph1 gene issufficient to prevent homoeologous pairing.

Additional gene loci that suppress homoeologous pairing are known in theart, as are mutations that relieve that control and thereby facilitatepairing and exchange between homoeologous chromosomes (see e.g., SearsE. R. (1976) Annual Review of Genetics 10:31-51). Although these variousgenes do not act in precisely the same fashion (see e.g., Martinez M.,Cuñado N., Carcelén N., Romero C. (2001) Theoretical and AppliedGenetics 103:398-405), any method known in the art that facilitatespairing of homoeologous chromosomes and homoeologous translocation ofgenetic material, specifically the transference of the Hardness locusfrom the short arm of chromosome 5D to either 5A or 5B or elsewhere inthe genome of tetraploid wheat where the gene locus will be stablyexpressed, is suitable for use in the construction of non-transgenictetraploid wheat plants having grain with soft kernel texture asdisclosed herein.

As is known in the art, homoeologous translocations are highlyunpredictable, variable, and are dependent on many factors such as e.g.,the particular chromosome involved, the varieties being used, the locusbeing transferred, the location of that locus, other undesirable genespresent on the translocated section of chromosome, differential gametetransmission rates, etc., and that T. turgidum is less tolerant ofgenome alterations than is T. aestivum (see e.g., Ceoloni C., Forte P.,Gennaro A. et al. (2005) Cytogenetic and Genomic Research 109:328-334;Luo M.-C., Yang Z.-L., Kota R. S., Dvo{hacek over (r)}ák J. (2000)Genetics 154:1301-1308; Jauhur P. P., Do{hacek over (g)}ramaci M. (2008)Euphytica 159:353-358; Juahur P. P., Almouslem A. B., Peterson T. S.,Joppa L. R. (1999) The Journal of Heredity 90:437-445). As will bediscussed in greater detail hereinbelow, the occurrence of both Ph1 andHardness on the same homoeologous chromosome group (group 5 chromosomes)presents an additional complicating factor in the possible cytogeneticmanipulation of these traits.

The methods disclosed herein for the construction of a non-transgenictetraploid wheat plants having grain with soft textured endospermutilize the variable presence or absence of chromosome 5B, the normallocation of the Ph1 gene locus. Ordinarily, a 5D(5B) disomicsubstitution line which lacks chromosome 5B would lack the Ph1 genewhich suppresses homoeologous pairing. Indeed, in the absence ofchromosome 5B there is some pairing with non-homologous chromosomes.Therefore, methods disclosed herein employ a substitution linecomprising a pair of 5D chromosomes and a monosome of 5B in order toprevent unwanted non-homologous pairing from occurring. The monosome isalmost never passed through the male parent, thus the substitution linewith the monosome 5B cannot participate in homoeologous pairing.

Although there have been some reports that allege successfulconstruction of non-transgenic tetraploid wheat plants having grain withsoft kernel texture, in each case it is apparent that the allegednon-transgenic tetraploid wheat plants having grain with soft kerneltexture do not behave as diploids in meiosis, e.g., they do not showstable Mendelian segregation. (see e.g., Gazza L., Niglio A., Mei E. etal. ((2002) Proceedings of second international workshop Durum Wheat andPasta Quality: Recent Achievements and New Trends, Rome, 19-20 Nov.2002, p. 285-288; Gazza L., Zanell L., and Pogna N. E. ((2008) 11^(th)International Wheat Genetics Symposium 2008, Proceedings of the 11^(th)International Wheat Genetics Symposium, 24-29 Aug. 2008, Brisbane,Queensland, Australia, vol. 2, p. 339-341; Simeone M. C., Lafiandra D.,Morris C. F. ((2003) Tenth International Wheat Genetics Symposium, 1-6Sep. 2003, Paestum, Italy, vol. 3, p. 1391-1393).

Thus, prior to the instant disclosure, non-transgenic tetraploid wheatplants having grain with soft kernel texture wherein the non-transgenictetraploid wheat plants having grain with soft kernel texture have afunctional Ph1 and diploid-like behavior in meiosis were not known, norhad a method for the construction of said wheat plants been heretoforeappreciated or disclosed.

C. “Herding Cats”—the Use of Ph1 Mutants Results in Loss of OrderedChromosome Pairing in Meiosis and Instability of the Wheat Genome

As is well known in the art, grass genomes are variably related based ontheir evolutionary relationships. In particular, the genomes ofhexaploid and tetraploid wheat are related to the degree that theirchromosomes are termed “homoeologues”. Homoeologues have, variably, theability to compensate for each other. Thus, through cytogeneticmanipulations, it is possible to construct disomic substitution lines intetraploid wheat using hexaploid wheat chromosomes.

Unfortunately, durum wheat exhibits a much lower tolerance toward genomealterations as compared to common wheat. Indeed, sizable alientransfers, even when well tolerated by the hexaploid common wheatgenome, are not well tolerated by the tetraploid durum wheat genome (seee.g., Ceoloni, C. et al. (2005) Cytogenetics and Plant Breeding109:328-334). For example, although tetraploid wheat plants having softtextured endosperm can be obtained by substituting the 5D chromosomesfrom hexaploid wheat for 5A or 5B in tetraploid wheat such disomicsubstitution lines are typically lower in grain yield and vigour and maybe sterile (see e.g., Liu et al. (1995) Journal of Cereal Science21:209-213).

Thus, whole chromosome substitutions or even Robertsonian whole armtranslocations are not typically desirable as parents in breeding stabletetraploid wheat cultivars because, 1) the alien chromosome or arm willnot pair during future breeding efforts e.g., an introduced 5D will havenothing to pair with nor will the 5A or 5B of the other parent dependingon the substitution, and 2) linkage drag—the association of undesirablegenes on the same chromosome or arm as the gene/trait of interest (seee.g., Qi, L. et al. (2007) Chromosome Research 15:3-19).

Linkage drag is thought to result from restrictions upon pairing betweenhomoeologous chromosomes. Thus, the transfer of a target gene from awild relative (often referred to as alien) to a crop plant is difficult,because transfer of the desired target gene is accompanied by thetransfer of other untargeted wild traits that are due to genes alsopresent in the transferred chromosome segment.

As is well known in the art, the problem of linkage drag andnon-acceptance of alien chromosome is typically overcome by removing thecontrolled pairing of homoeologues exerted by the Ph1 gene locus. ThePh1 can be removed by removing the entire 5B chromosome, removing thelong arm of 5B, or deleting some smaller portion of 5BL via irradiation.Two exemplary radiation-induced mutants are the hexaploid wheat ph1b andthe tetraploid wheat ph1c. In wheat plants lacking Ph1 control,homoeologues may pair, form chiasmata and exchange genetic material.

Movement of traits from one genome to another by removing Ph1 control isreferred to as “homoeologous translocation.” Typically, at some pointafter homoeologous translocation is effected, Ph1-mediated control mustbe restored by replacing the mutation with the functional gene locus soas to stabilize the newly created line.

Indeed, it is well known in the art that the use of Ph1 mutants ishighly unpredictable (see e.g., Qi, L. et al. (2007) supra; Sears, E. R.(1977) Canadian Journal of Genetics and Cytology 19:585-593). The lackof Ph1 control permits homeologous pairing and recombination, which canvary between different chromosomes of a species, and between the shortand long arms of a chromosome. Furthermore, wheat plants lacking Ph1control experience a high frequency of recombination betweennon-designated homoeologues as well as non-homologous pairing. Thus, theproducts of meiosis and the progeny resulting therefrom are highlyunpredictable (see e.g., Qi, L. et al. (2007) supra, Jauhar, P. P. andDogramaci, M. (2008) Euphytica 159:353-358). Thus, Ph1 mutant parentlines are inherently unstable and progeny of Ph1 mutant parentstypically suffer chaotic chromosome rearrangements. Indeed, as a resultof the irregularities associated with homeologous pairing, progeny ofPh1 mutant parents are known to carry monosomes and trisomes at highfrequency, as well as telocentric chromosomes and other chromosomeirregularities (see e.g. Sears, E. R. (1977) supra).

Since the Ph1 locus is on the 5B chromosome, attempts at homoeologoustranslocations involving group 5 chromosomes are especiallyunpredictable. Although this general approach (homoeologoustranslocation) has proven successful in tetraploid wheat, a much greaterand further complication results when the gene/trait of interest lies inone of the homoeologous group 5 chromosomes as does the gene conferringsoft textured kernels which is located on chromosome 5D (see e.g.,Joppa, L. R., and Williams, N. D. (1983) “Genetics and breeding of durumwheat in the United States.” In Durum Wheat: Chemistry and Technology.Eds. G. Fabriani and C. Lintas, American Association of Cereal Chemists,Inc., St. Paul, Minn., Chapter 3, pp. 47-67; Sears, E. R. (1976) AnnualReviews Genetics 10:31-51; Sears, E. R. (1977) supra).

Since chromosomal rearrangements, unbalanced genomic constitutions,aneuploidies, structural and intergenomic rearrangements are commonplacein wheat plants lacking Ph1 control (see e.g., Sánchez-Moran et al.(2001) Chromosoma 110:371-377) and since both the Ph1 locus and thesoftness locus are located on the group 5 chromosomes, predicting theoutcome from crosses employing a homeologous translocation of the 5Dchromosome into tetraploid wheat is so to speak, tantamount to “herdingcats”.

D. Construction of a Non-Transgenic Tetraploid Wheat Plant Having SoftTextured Endosperm Using Classical Breeding Techniques

It has now been discovered that teraploid wheat plants that producewheat grain having a soft texture (soft textured endosperm) can beconstructed using classical and cytogenetic breeding techniques. In anexemplary embodiment, a non-transgenic tetraploid wheat plant havingsoft textured endosperm is a durum wheat plant.

In general, the goal of wheat breeding is to develop new, unique andsuperior wheat varieties. In practical application of a wheat breedingprogram, a breeder initially selects and crosses two or more parentallines, followed by repeated selfing and selection, producing many newgenetic combinations. Theoretically billions of different geneticcombinations can be generated via crossing, selfing and mutations. Anexemplary method for constructing a non-transgenic durum wheat planthaving soft textured endosperm is disclosed hereinbelow.

ABBREVIATIONS/DEFINITIONS

-   -   LDN=Langdon    -   CS=Chinese Spring    -   CS-5D(5B)=CS carrying a disomic substitution where 5D have        replaced 5B    -   CS(ph1b)=CS carrying the ph1b mutation    -   LDN-ddt-5B=LDN double ditelosomic 5B    -   ph1b=Pairing homoeologous-1 gene with deletion (=b allele)

Stocks:

-   -   LDN-47-1 with ph1b from CS (maintained as an F₁ heterozygote)    -   LDN-16    -   LDN double ditelosomic 5B    -   LDN-5D(5B) substitution line

Pedigrees:

-   -   LDN-5D(5B) substitution line: CS-5D(5B)/*12 LDN-16    -   LDN-47-1 ph1b line: CS(ph1b)/*2 LDN-16//2*LDN-47-1 [F₂ seed,        maintained as a heterozygote]

Step 1:

Cross ‘LDN-5D(5B)’×‘LDN-47-1’ (disomic substitution line with the ph1bmutant line).

Results:

F₁ plant(s) that is(are) monosomic for both 5D and 5B; some of theprogeny will also have ph1b.

Step 2:

Grow F1 plants, select plants with 2n=28 chromosomes. Those with 13bivalents+2 monovalents do not carry ph1b, whereas those with variablepairing do carry ph1b; self-fertilize; those plants with ph1b will havehomoeologous pairing between 5D and 5B and crossing-over (creatingtranslocation(s)).

Step 3:

Harvest F₂ seeds; select plants with variable pairing that carry ph1b.In some cases, at least some of the plants will have cells in which atleast one of the chromosomes will fail to form ring bivalents. This mayindicate that one of the chromosomes is a translocation between 5B and5D.

Step 4:

Cross selected F₂ plants to a “normal durum” (for example, crossing toLangdon represents BC₁).

Step 5:

Select BC₁F₁ plants with 13 pairs of chromosomes at metaphase I ofmeiosis and variable pairing; self fertilize.

Step 6:

Grow BC₁F₂ plants.

Step 7a: ‘Test’ Cross (verify translocation vs. a 5D disomicsubstitution)

Cross BC₁F₂ plants with LDN-ddt-5B.

Step 7b:

Grow progeny. At least three types of progeny are expected. If theparent had a pair of 5D chromosomes (a substitution, not atranslocation), then the 5B telosomic chromosomes will fail to pair. Ifthe parent had a pair of 5B chromosomes, then both telosomic chromosomeswill pair. If the parent had a translocation, then only one of thetelosomic chromosomes will pair (the 5BL).

Step 7c:

Select plants with 13 bivalents=1 telomeric bivalent+1 telomericmonovalent (plants with translocation). Discard plants with 13bivalents+trivalent with one whole chromosome and two telosomicchromosomes, one for each arm all paired together, in short handnotation, tlt′″, i.e., a 5D disomic substitution).

Step 8:

Cross to LDN-5D(5B) substitution line and look for the parents in whichonly one of the chromosomes fails to form ring bivalents. Select parentsbased on this cytological analysis; grow and self-fertilize.

The above protocol shows exemplary steps taken to construct anon-transgenic tetraploid wheat plant having grain with soft kernaltexture. As will be appreciated by a person having skill in the art,other durum wheats other than Langdon, can be used (if available). Aperson of skill in the art also well appreciates that ph1c or othermeans to relieve homoeologous pairing restrictions could be used.Disomic substitutions other than Chinese Spring are also suitable,provided the 5D carries to the Hardness locus with expressed Pina andPinb. As will be readily appreciated by a person of skill in the art,the key is the “composition” and nature of the stocks and how they areused sequentially, not the exact variety background in which they exist.

III. Measuring Endosperm Texture

Grain texture classification is based primarily on either the resistanceof kernels to crushing or the particle size distribution of ground grainor flour. Because kernel texture is important for wheat grain qualityand utilization, numerous methods have been developed to measure graintexture (see e.g., Pomeranz, Y. and Williams, P. C. (1990) Wheathardness: Its genetic, structural, and biochemical background,measurement, and significance. Pages 471-548 In: Advances in CerealScience and Technology, Vol. X. Y. Pomeranz, ed. AACC International: St.Paul, Minn.; Glenn, G. M. et al. (1991) J. Cereal Sci. 13:179-194;Haddad et al. (1998) Cereal Chem. 75:673-676; Morris (2002) Plant Mol.Biol. 48:633-347; Morris, C. F. et al. (2007) Cereal Chem. 84:67-69;Morris, C. F. et al. (2008) Cereal Chem. 85:351-358; Pearson, T. et al.(2007) Cereal Chem. 84:567-575.

Typically, methods for measuring grain hardness measure either: (1) thesize and distribution of particles after grinding or milling, or (2)directly measure the compressive strength of kernels.

Exemplary methods for measuring the size and distribution of particlesafter grinding or milling include, but are not limited to the particlesize index (PSI) (see e.g., Williams, P. C. and Sobering, D. C. (1986)Cereal Foods World 31:359, 362-364 (Approved Method 55-30, AACCInternational 2000); and near infra-red (NIR) spectroscopy (see e.g.,Norris, K. H. et al. (1989) Cereal Foods World 34:696-705 (ApprovedMethod 39-70A, AACC International 2000).

Exemplary methods for directly measuring the compressive strength ofkernels include, but are not limited to automated weighing and crushingof individual kernels (see e.g., Martin, C. R. et al. (1993) Trans. ASAE36:1399-1404). Instruments capable of directly weighing and crushingIndividual kernels are available and known to those of skill in the art.For example, the single kernel characterization system (SKCS) model 4100(Perten Instruments, Springfield, Ill.) (AACC Approved Method 55-31) iseffective for carrying out measurement of the compressive strength ofkernels.

In an exemplary embodiment, wheat grain having soft textured endospermis measured according to the Perten SKCS 4100 (Perten Instruments, Reno,Nev. See e.g., Instruction Manual, Single-Kernel CharacterizationSystem, Model SKCS 4100; Perten Instruments, Inc.: 6444 South SixthStreet Road, Springfield, Ill. 62707) and has a hardness index/value ofbetween about 10 to about 45 on a scale of −20 to 120. In otherexemplary embodiments, the kernel hardness associated with grain of softwheats have SKCS hardness values in a range that is from about 0 toabout 55. In other exemplary embodiments, the grain of a non-transgenictetraploid wheat plant having soft textured endosperm have SKCS hardnessvalues in a range that is from about 15 to about 35. In still otherexemplary embodiments, the grain of a non-transgenic tetraploid wheatplant having soft textured endosperm has a hardness value that is in arange that is between about 10 to about 45 as measured by the PertenSKCS 4100.

IV. Preparing Semolina, Flour and Pasta

In exemplary embodiments, soft textured grain produced by anon-transgenic tetraploid wheat plant as disclosed herein, is milled toprepare durum flour and/or semolina.

Wheat grain is rarely consumed whole and as such, is first converted togranular products via milling. Wheat grain milling is largely dependenton kernel texture and thus, a mill is typically designed to millexclusively soft wheat, hard wheat or durum wheat, in some instances amill may be a “swing” mill such that it can, at lower efficiency, millboth soft and hard wheats.

Typically durum wheat is milled to produce a product of coarsegranulation called semolina (see e.g., CFR §137.320); the production ofdurum flour (see e.g., CFR §137.220) is not a primary objective butcannot be wholly prevented as some few grain particles will be ofinsufficiently small size to be construed as flour. Pasta manufacturersprefer finer granulations and durum flour receives little if anymonetary discount in the trade. The primary obstacle to producing agreater proportion of durum flour relative to semolina resides in thegreater work involved in obtaining the finer granulation and theinherent greater level of damaged starch as work input increases (seee.g., Posner, E. S., and Hibbs, A. N. (1997) Wheat Flour Milling. St.Paul, Minn.: American Association of Cereal Chemists, Inc.).

Methods for preparing semolina are known in the art (see e.g., U.S. Pat.No. 5,141,764; U.S. Pat. No. 7,506,829; Approved Method 26-42, AACCInternational 2000).

In an exemplary embodiment, durum wheat is milled to produce coarse,granular particles of semolina. Typically, in the production of durumsemolina, it is desirable to minimize the production of durum flour.Therefore, the primary difference between flour and semolina milling isthe method and degree of grinding of the endosperm. In general flour isfinely ground, whereas semolina is coarsely ground.

In an exemplary embodiment, the semolina prepared from soft texturedendosperm produced by a non-transgenic tetraploid wheat plant asdisclosed herein, is used for making pasta. Pasta manufacturing is knownin the art (see e.g., U.S. Pat. No. 6,326,049; U.S. Pat. No. 3,520,702;U.S. Pat. No. 6,322,840; U.S. Pat. No. 6,203,840).

As is known in the art, pasta manufacturers typically prefer themajority of particles to fall within a narrow particle size range toinsure that semolina will flow freely, and that pasta dough water uptakewill be homogeneous (see e.g., Feillet, P. and Dexter, J. (1996) Qualityrequirements of durum wheat for semolina milling and pasta production.Pp. 95-131 In: Pasta and Noodle Technology. Kruger, J., Matsuo, R. andDick, J., editors, American Association of Cereal Chemists. St. Paul,Minn.).

In an exemplary embodiment grain from a non-transgenic tetraploid wheatplant having grain with soft kernel texture is milled to flour whereinthe resulting flour has low levels of damaged starch. In anotherexemplary embodiment, grain from a non-transgenic tetraploid wheat planthaving grain with soft kernel texture is milled to produce durumsemolina with low work expenditure.

V. Measuring Starch Damage

Some starch granules are mechanically damaged during the millingprocess. Damage is typically manifest as physical cracks and breaks inthe starch granule. Since the level of starch damage directly affectswater absorption, dough mixing properties and other qualities of themilled product e.g., semolina, flour, etc., starch damage is oftechnological significance. Thus, it is useful to be able tocharacterize starch damage in terms of both quantity and quality.

In general, the greater the compressive and shearing forces to achieveda granular milled product e.g., semolina, flour, etc, the more starchdamage may be incurred. Thus, typically harder textured grain whichrequires the application of greater compressive and shearing forces toachieve a milled granular product, typically experiences greater starchdamage during milling than a softer textured grain.

Methods for measuring starch damage are known in the art. Most methodscan be grouped in general, into four classes: extraction procedures(“Blue Value”), dye-staining procedures, NIR procedures, and enzymedigestion procedures (see e.g., Williams, P., and Fegol, K. (1969)Cereal Chemistry 46, 56-6; Chiang, B., et al. (1973) Cereal chemistry50:44-49; Medcalf, D. and Gilles, K. (1965) Cereal Chemistry 42:546-557.A. D. Evers, and D. J. Stevens (2006) Starch 40(8):297-299; Evers, A. D.and Stevens, D. J. (1985). “Starch Damage” In: “Advances in CerealScience and Technology” Vol. VII. (Pomeranz, Y. Ed.) AmericanAssociation of Cereal Chemists Inc. St. Paul Minn., pp. 321-349; Gibson,T. S., Al Qalla, H. and McCleary, B. V. (1991) J. Cereal Sci., 15,15-27; Gibson, T. S., Kaldor, C. J. and McCleary, B. V. (1993) CerealChem., 70:47-51; American Association of Cereal Chemists (AACC) ApprovedMethod 76-31).

Any method known in the art for measuring starch damage is suitable foruse in the methods disclosed herein. A person having skill in the artand access to the instant specification will appreciate and choose themethod best suited to their circumstances for the purpose of determiningstarch damage.

In one exemplary embodiment, starch damage is determined using AACCMethod 76-31 Damaged Starch: Spectrophotometric Method (see e.g.,Approved Methods of the American Association of Cereal Chemists (AACC),10th Edition, 2003) and starch damage is expressed as a percentage offlour weight. AACC Method 76-31 is sometimes referred to as the Megazymemethod.

In another exemplary embodiment, starch damage is determined using AACCMethod 76-30A Damaged Starch.

VI. Deposit Information

A deposit of the non-transgenic tetraploid durum wheat strain havingsoft textured endosperm, Soft Svevo Durum Wheat WAS 080240001, disclosedhereinbelow and recited in the appended claims has been made with theAmerican Type Culture Collection (ATCC), Patent Depository, 10801University Blvd., Manassas, Va. 20110, U.S.A. The date of deposit wasMay 27, 2009. All restrictions upon the deposit have been removed, andthe deposit is intended to meet all of the requirements of 37 C.F.R.1.801-1.809. The ATCC accession number is PTA-10087. The materialdescription is: Durum Wheat from Italy, Soft Svevo durum wheat WAS080240001. The deposit will be maintained in the depository for a periodof 30 years, or 5 years after the last request, or for the effectivelife of the patent, whichever is longer, and will be replaced asnecessary during that period.

The following examples are offered to illustrate, but not to limit theinvention.

EXAMPLES Example 1

The following example illustrates construction and testing of anon-transgenic tetraploid wheat plant having soft textured endospermusing classical genetic techniques. In particular the following Exampleillustrates construction of the line known as: Soft Svevo Durum Wheat,WAS 080240001, which was deposited with the American type CultureCollection on May 27, 2009 and which has been assigned ATCC accessionnumber is PTA-10087.

Fourteen (14) putative non-transgenic soft textured durum wheat grainvarieties, bearing a putative homoeologous translocation were produced.Six of these being identified as ‘674’, ‘675’, ‘678’, ‘679’, ‘685’, and‘688’, were used as disclosed hereinbelow. Each of the six abovereferenced putative non-transgenic soft textured durum wheat grainvarieties was crossed to the durum wheat cultivars Svevo and Creso, andthe progeny of each was evaluated for soft kernel texture using thePerten SKCS 4100. Results are provided in FIGS. 2-13 and hereinbelow.

ABBREVIATIONS/DEFINITIONS

-   -   LDN=Langdon    -   CS=Chinese Spring    -   CS-5D(5B)=CS carrying a disomic substitution where 5D have        replaced 5B (same as CS Nullisomic 5B/Tetrasomic 5D)    -   CS(ph1b)=CS carrying the ph1b mutation (the ph1b mutation allows        homeologous pairing)    -   LDN-dbl ditelo-5B=LDN double ditelosomic 5B    -   LDN-47-1=LDN with ph1b from CS (maintained as an F₁ heterozygote        due to the presence of ph1b mutation)    -   ph1b=Pairing homoeologous-1 gene with deletion (=b allele)

Stocks:

LDN-47-1

LDN-16

LDN dbl ditelo-5B

LDN-5D(5B) disomic substitution line. The LDN-5D(5B) line is preferablymaintained with a 5B monosome and is used as male in crosses. Normally,a 2n=28 disomic addition line Langdon (LDN)-5D(5B) would have no 5Bchromosome, and therefore no functional Ph1, thus its genome is“unstable” each time the genome experiences meiosis. This phenomenon isdiscussed e.g., in Section II C. hereinabove “Herding Cats”. Thereforeit is preferably maintained with a 5B monosome (2n=29). Use of theLangdon (LDN)-5D(5B) strain with a 5B monosome as a male in crossesensures that the monosome is not transmitted to the progeny since themonosome is not transmitted in the pollen. Thus, maintenance ofLDN-5D(5B) with a 5B monosome and its use as the male in crosses ensuressuccess in creating the non-transgenic tetraploid wheat plants havinggrain with soft textured endosperm as disclosed herein.

Pedigrees:

LDN-5D(5B) disomic substitution line: CS-5D(5B)/*12 LDN-16

LDN-47-1 ph1b line: CS(ph1b)/*2 LDN-16//2*LDN-47-1 [F₂ seed, maintainedas a heterozygote, which will facilitates construction of thenon-transgenic tetraploid wheat plants having grain with soft texturedendosperm as disclosed herein.]

An exemplary series of crosses used to produce non-transgenic tetraploidwheat plants having soft textured endosperm is disclosed hereinbelow:

Step 1:

‘LDN-5D(5B)’×‘LDN-47-1’ were crossed (i.e., disomic substitution linecrossed with the ph1b mutant line).

The resulting F₁ plants were monosomic for both 5D and 5B; some of theprogeny also carried ph1b.

Step 2:

The F1 plants produced in Step 1, were grown and plants with 2n=28chromosomes were selected by methods known in the art. Common techniquesinclude, but are not limited to e.g., preparing root tip squashes withstaining and visualization of chromosomes (see e.g. R. J. Singh, (2003)Plant Cytogenetics, 2^(nd) Edition, CRS Press, Boca Raton). Plants with13 bivalents+2 monovalents do not carry ph1b, whereas those withvariable pairing do carry ph1b. Plants were self-fertilized. Thoseplants with ph1b experience homoeologous pairing between 5D and 5B andcrossing-over (creating translocation(s)).

Step 3:

F₂ seeds were harvested and plants with variable pairing which thereforecarry ph1b were selected. In some cases, at least some of the plants hadcells in which at least one of the chromosomes will fail to form ringbivalents (see e.g., N. S. Cohn, 1969, Elements of Cytology, 2^(nd)Edition, Harcourt, Brace & World, Inc., New York; R. J. Singh (2003)supra) suggesting that one of the chromosomes was a translocationbetween 5B and 5D.

Step 4:

The selected F₂ plants carrying ph1b, were crossed to a Langdon durum(represents BC₁).

Step 5:

BC₁F₁ plants with 13 pairs of chromosomes at metaphase I of meiosis andvariable pairing were selected using methods known in the art (see e.g.,R. J. Singh (2003) supra; K. S. Quisenberry and L. P. Reitz (1967)supra) and the selected plants were self fertilized.

Step 6:

F2 plants from the self fertilization in step 5 i.e., BC₁F₂ plants, weregrown to maturity.

Step 7a:

“Test Crosses” were conducted to determine if the selected BC₁F₂ plantcarried a translocation vs. a 5D disomic substitution.

Thus, BC₁F₂ plants were crossed with LDN dbl ditelo-5B.

As is appreciated by a person of skill in the art, If the entire 5D isthere (disomic substitution, then both the long arm and short armditelos will fail to pair with it, in the ‘squash’ under the microscopeone sees the two arms and the 5D monosome—none paired. If it was a 5Bdisomic substitution (whole 5B present), then both the 5BS and 5BLditelos can pair with the 5B whole. 5BS/5DS translocations (withHardness), then 5BL will pair with the translocated chromosome (the longarm of 5B should still be intact), BUT the presence of the 5DS bit willprevent the 5BL ditelo from pairing.

Step 7b:

Progeny from the test cross in 7a were grown to maturity. At least threetypes of progeny are expected. As is appreciated by a person of skill inthe art, If the entire 5D is there (disomic substitution, then both thelong arm and short arm ditelos will fail to pair with it, in the‘squash’ under the microscope one sees the two arms and the 5Dmonosome—none paired. If it was a 5B disomic substitution (whole 5Bpresent), then both the 5BS and 5BL ditelos can pair with the 5B whole.5BS/5DS translocations (with Hardness), then 5BL will pair with thetranslocated chromosome (the long arm of 5B should still be intact), BUTthe presence of the 5DS bit will prevent the 5BL ditelo from pairing. Ingeneral, for reasons discussed in detail herein above, see Section II.C. “Herding cats”, progeny having a whole pair of 5D chromosomes or awhole-arm of 5D chromosomes are considered unsuitable for breeding.

Step 7c:

Plants with 13 bivalents=1 telomeric bivalent+1 telomeric monovalent(i.e., the plants with a translocation) were selected. Plants with 13bivalents+trivalent with one whole chromosome and two telosomicchromosomes, one for each arm all paired together, in short handnotation, tlt′″, i.e., a 5D disomic substitution were discarded.

Step 8:

The plants selected in step 7c, were crossed to LDN-5D(5B) disomicsubstitution line and were examined to determine the parents in whichonly one of the chromosomes failed to form ring bivalents usingcytological methods known in the art (see e.g., R. J. Singh (2003)supra). The parents in which only one of the chromosomes failed to formring bivalents were selected on the cytological analysis. Selectedplants were grown to maturity and were self-fertilized. F3:4 seeds wereharvested.

Step 9:

We obtained a total of 438 F_(3:4) seeds from 14 F₃ experimental lines.Approximately half of the seeds of each line (total=170 seeds) were usedto grow plants in the greenhouse. Approximately half the plants weresterile and a significant proportion died of low vigor. Some plants, onthe other hand, were highly vigorous. Vigor was independent from the F₃line origin. These plants were used as pollinators in crossing to theItalian durum cultivars Svevo and Creso. Successful hybridizationsproduced 308 F₁ kernels (132 from Creso, 176 from Svevo).

The F3:4 progeny were grown to maturity, self fertilized, and the F3:5seed was harvested. This process produced 14 putative Langdon durumhomoeologous translocation lines, six of which are referred to herein as‘674’, ‘675’, ‘678’, ‘679’, ‘685’, and ‘688’ were used in crosses toCreso and Svevo.

Step 10:

The same F3:4 progeny were grown and were “Test Crossed” to Svevo andCreso durum cultivars. F1 seed was harvested and F1 plants were grown tomaturity and self fertilized. F2 seed was harvested on a per plantbasis; and kernel texture phenotype was assessed. Specifics of the “TestCrosses” are discussed below.

F3:5 non-vitreous appearing kernels of Langdon durum homoeologoustranslocation lines 678 and 683, ten plants each, were increased in agreenhouse to produce F6 seed. Seed from the ten plants of each of thetwo lines was bulked and evaluated for SKCS kernel texture. The resultwas Hardness Indexes of 34.4±30.5 for 678 and 41.0±32.8 for 683,indicating that both were likely heterogeneous for soft and hardkernels, based on the moderately low means and markedly high standarddeviations.

Langdon durum homoeologous putative translocation line 674 was crossedto Svevo durum cultivar and produced progeny with SKCS kernel texturefrom 5 to 80 (FIG. 2). 80 (FIG. 2). Segregation was unpredictable andhighly chaotic but recovery of soft progeny indicated that line 674possessed a homoeologous translocation bearing the Hardness locus.Phenotypic segregation was not predictable and did not conform to anyexpected genetic ratio, indicating potential genome instability.

Langdon durum homoeologous translocation line 674 was crossed to Cresodurum cultivar and produced progeny with SKCS kernel texture from 22 to83 (FIG. 3); phenotypic segregation was not predictable and did notconform to any expected genetic ratio, indicating potential genomeinstability.

Langdon durum homoeologous translocation line 675 was crossed to Svevodurum cultivar and produced progeny with SKCS kernel texture from 17 to82 (FIG. 4); phenotypic segregation was not predictable and did notconform to any expected genetic ratio, indicating potential genomeinstability.

Langdon durum homoeologous translocation line 675 was crossed to Cresodurum cultivar and produced progeny with SKCS kernel texture from 38 to57 (FIG. 5); phenotypic segregation was not predictable and did notconform to any expected genetic ratio, indicating potential genomeinstability.

Langdon durum homoeologous translocation line 678 was crossed to Svevodurum cultivar and produced progeny with SKCS kernel texture from 27 to76 (FIG. 6); phenotypic segregation was not predictable and did notconform to any expected genetic ratio, indicating potential genomeinstability.

Langdon durum homoeologous translocation line 678 was crossed to Cresodurum cultivar and produced progeny with SKCS kernel texture from 23 to63 (FIG. 7); phenotypic segregation was not predictable and did notconform to any expected genetic ratio, indicating potential genomeinstability.

Langdon durum homoeologous translocation line 679 was crossed to Svevodurum cultivar and produced progeny with SKCS kernel texture from 57 to77 (FIG. 8); phenotypic segregation was not predictable and did notconform to any expected genetic ratio, indicating potential genomeinstability.

Langdon durum homoeologous translocation line 679 was crossed to Cresodurum cultivar and produced progeny with SKCS kernel texture from 50 to80 (FIG. 9); phenotypic segregation was not predictable and did notconform to any expected genetic ratio, indicating potential genomeinstability.

Langdon durum homoeologous translocation line 685 was crossed to Svevodurum cultivar and produced progeny with SKCS kernel texture from 28 to63 (FIG. 10); phenotypic segregation was not predictable and did notconform to any expected genetic ratio, indicating potential genomeinstability.

Langdon durum homoeologous translocation line 685 was crossed to Cresodurum cultivar and produced progeny with SKCS kernel texture from 22 to82 (FIG. 11); phenotypic segregation was not predictable and did notconform to any expected genetic ratio, indicating potential genomeinstability.

Langdon durum homoeologous translocation line 688 was crossed to Svevodurum cultivar and produced progeny with SKCS kernel texture from 12 to81 (FIG. 12); phenotypic segregation was not predictable and did notconform to any expected genetic ratio, indicating potential genomeinstability.

Langdon durum homoeologous translocation line 688 was crossed to Cresodurum cultivar and produced progeny with SKCS kernel texture from 18 to78 (FIG. 13); phenotypic segregation was not predictable and did notconform to any expected genetic ratio, indicating potential genomeinstability.

As demonstrated above, all six Langdon durum homoeologous translocationlines that were produced as disclosed in steps 1-9 of this examplecomprised the homoeologous translocation carrying the Hardness genelocus, in either the homozygous or heterozygous condition. Also clear isthat all the lines were segregating and/or were unstable i.e., did notbehave as diploids in meiosis. As will be demonstrated hereinbelow, thetranslocations were crossed, selected and stabilized to providenon-transgenic durum wheat plants having grain with soft kernel texture.

Seeds from each of 10 plants from each of lines 678 and 683 were bulkedand subjected to SKCS kernel texture analysis. Results showed them to behighly heterogeneous for kernel texture. Mean hardness index andstandard deviation were 34±31 and 41±33, line 678 and 683, respectively.The SKCS also produces a four-class histogram with hardness index limitsof #33, 34-46, 47-59, and 960. For line 678 the percentages of kernelsin each class were 59, 5, 6 and 30, and for line 683 the percentageswere 48, 7, 7 and 38. These data clearly indicated that as a population,the kernel texture distributions for both lines were highly bimodal withabout half the kernels markedly soft (i.e. Hardness Index #33).

F2 seeds from a small sub-sample of the F1 plants of the crossesCreso/685 and Creso/688 described above were grown, self fertilized andassayed for SKCS kernel texture. One plant each of crosses Creso/685 andCreso/688 produced Hardness Index and standard deviation of 69±24 and29±8, respectively. Five plants from the Svevo/675 cross producedhardness index and standard deviation of 38±10, 56±19, 32±18, 22±20 and19±20. One to five F3 plants from each line were grown to maturity,harvested and the seed used to plant F3:4 rows at the Washington StateUniversity Spillman Agronomy Farm, Pullman, Wash. Thirty-four rows intotal were harvested and the F5 seed assayed for SKCS kernel texture(FIG. 14). The distribution of kernel texture phenotype was in keepingwith a single locus-two allele model wherein those lines with hardnessindex greater than 65 and standard deviation less than 16 putativelylacked the Hardness-containing translocation and were uniformly hard,those with hardness index from about 40 to 65 and standard deviationgreater than 18.3 were putatively heterogeneous, and those with hardnessindex less than 35 were soft and putatively carried theHardness-containing translocation. In this group of soft wheat lines,the standard deviation ranged from 11.2-18.4. The two durum parents,Creso and Svevo, were located in the ‘hard’ group.

As disclosed below, crosses between Langdon durum homoeologoustranslocation line 674 and Svevo were conducted to provide anon-transgenic soft durum wheat variety referred to herein as Soft Svevodurum wheat WAS 080240001, representative seed of such line having beendeposited under ATCC Accession No. PTA-10087.

Line 152 was selected from the cross between Langdon durum homoeologoustranslocation line 674 and Svevo (see FIG. 2). Although one line wasselected other soft kernel progeny from this cross as well as progenyfrom any and all other crosses would have been expected to producedsimilar results. This particular line was selected as it exhibited theoverall softest kernel texture and therefore was expected to producesoft progeny with stable soft kernel inheritance. DNA was extracted fromdistal half seeds (n=10) and assayed using PCR and puroindoline a and bprimers. To amplify puroindoline-a gene(s) the sense-strand primer was5′-ATGAAGGCCCTCTTCCTCA-3′ (SEQ ID NO: 1) and the antisense-strand primerwas 5′-TCACCAGTAATAGCCAATAGTG-3′ (SEQ ID NO:2). To amplifypuroindoline-b gene(s) the sense-strand primer was5′-ATGAAGACCTTATTCCTCCTA-3′ (SEQ ID NO:3) and the antisense-strandprimer was 5′-TCACCAGTAATAGCCACTAGGGAA-3′ (SEQ ID NO:4). PCR was carriedout by known methods (see e.g., Sambrook and Russell (2001) MolecularCloning, A Laboratory Manual, CSH Press).

Six of the ten seeds were positive for both puroindoline a and b. Theremaining embryo half of these seeds were germinated and the plantsgrown, emasculated and used for crossing to Svevo (as female)(representing back-cross 1, BC1). Resultant progeny were assayed forpuroindoline genes via PCR as before (ten of 19 seeds were positive forboth genes). PCR-positive individual seeds were propagated and used forBC2. The process was repeated for BC3. Puroindoline PCR-positive progenywere grown in the field near Viterbo, Italy, and allowed to selfpollinate. Four to seven BC3F2 seeds from each of these BC3F1 plantswere subjected to SKCS kernel texture analysis on an individual plantbasis (FIG. 15). A total of 92 plants were evaluated along with twoplants of Svevo. From these results, a single plant (identified as no.88) was selected as having a high probability of being uniformly softand carrying the Hardness translocation. This selection was advancedthrough two more cycles of self pollination to produce BC3F4 seed.

A 500-gram aliquot of BC3F4 seed of line no. 88 was provided to Dr. KimShantz, WestBred LLC, Yuma, Ariz., for field increase during the2007-2008 crop season. Approximately 135 kg of BC3F5 seed was harvestedin 2008. The kernel texture of this field increase was hardness index24±14, indicating that it was uniformly soft. Dr. Shantz, described lineno. 88 as being uniform and otherwise indistinguishable from Svevo. Lineno. 88 was herein designated Soft Svevo durum wheat WAS 080240001,representative seed from this Arizona production having been depositedunder ATCC Accession No. PTA-10087.

Example 2 Deposit Information

Representative of, but not limiting the invention, Applicants havedeposited seeds from Soft Svevo durum wheat WAS 080240001, with theAmerican Type Culture Collection.

Applicants have made available to the public without restriction adeposit of at least 2500 seeds of a non-transgenic tetraploid wheatplant having soft textured endosperm i.e., Soft Svevo durum wheat WAS080240001, with the American Type Culture Collection (ATCC), Rockville,Md. 20852. The deposit was made May 27, 2009, under ATCC Accession No.PTA-10087.

The deposit will be maintained in the ATCC depository, which is a publicdepository, for a period of 30 years, or 5 years after the most recentrequest, or for the effective life of the patent, whichever is longer,and will be replaced if a deposit becomes nonviable during that period.

Example 3

The following example illustrates use of the non-transgenic putativesoft kernel Langdon durum homoeologous translocation line 678 in a crossto Kyle durum cultivar and the production of progeny with SKCS kerneltexture from 49 to 96 (FIG. 16). The non-transgenic putative soft kernelLangdon durum homoeologous translocation line 683 was crossed to Kyledurum cultivar and produced progeny with SKCS kernel texture from 48 to95 (FIG. 17).

One F1 plant of the pedigree Kyle/683 with a hardness index of 49±27(identified in FIGS. 16 and 17) was selected, grown, and its progenytested for kernel texture segregation. Nineteen F2 plants were grown tomaturity, harvested, and the F3 kernels subjected to SKCS kernel textureanalysis (FIG. 18). Most F2 lines could be classified based on kerneltexture phenotype: one line was soft with a hardness index of 16±17, and3-5 lines were hard with hardness indexes greater than 81. However,three of these had relatively low standard deviations (<17) whereas theother two had high standard deviations (>21). The remainder had hardnessindexes from 47 to 70; all but one had standard deviations greater than22. The segregation ratio indicated that the soft class may have beenunder-represented.

Two F2 plants of the cross Kyle/678 (those with hardness indexes of57±27 and 59±23) and three of the cross Kyle/683 (those with hardnessindexes of 49±27, 51±31 and 57±27) were selected and advanced(identified in FIGS. 16 and 17). F2 plants were grown and back-crossedto Kyle. BC1F1 plants were grown in a glasshouse, harvested and the seedfrom eight plants from each cross was subjected to SKCS kernel textureanalysis (FIG. 19). Mean hardness index ranged from 35 to 78, and allplants had hardness index standard deviation of 19 or greater. Progenyresulting from the Kyle/678 cross were more variable in hardness indexthan were those from Kyle/683. A soft line was clearly recovered fromthe Kyle/678 cross (hardness index 35). Setting delineations at about 37and 67 hardness index, the progeny of Kyle/678 approximated a 1:2:1segregation ratio, whereas the progeny of the Kyle/683 cross all fellinto the intermediate class of kernel texture.

Example 4

The following example illustrates the use of the non-transgenic SoftSvevo durum wheat WAS 080240001, representative seed having beendeposited under ATCC Accession No. PTA-10087, was used as a parent toproduce a non-transgenic soft textured durum wheat grain cultivar.

BC3F4 plants of Soft Svevo durum wheat WAS 080240001 were grown aspollen donor for crossing to Havasu durum wheat cultivar (female). F1seed was harvested and nineteen F1 plants were grown to maturity,harvested and the seed subjected to SKCS kernel texture analysis (FIG.20). The results were consistent with a single gene, bi-allelicsegregation model with a mid-range of kernel texture from hardness indexof 41 to 62, all with standard deviations greater than 21. Inspection ofthe SKCS four-class histogram results showed strong bi-modality for allF1 plants where the greatest proportion of kernels of each line were inthe #33 and 960 classes consistent with normal F2 segregation withineach spike. Seven F1 plants were selected and about five F2 plants ofeach were grown and crossed “blind” to Havasu. About half the crosseswere successful; non-emasculated F2 spikes were allowed to selfpollinate to produce F3 seeds. These F3 seeds were used as a ‘progenytest’ to evaluate the genotype of the parental F2 plants.

The F2 plants showed a characteristic normal one gene locus segregation,wherein a distinct three-cluster pattern was observed: a group of low(soft) hardness index with standard deviation less than 14, a group ofhigh (hard) hardness index with standard deviation less than 16, and anintermediate hardness index group with hardness index standard deviationgreater than 25 indicating ‘mixed’ (segregating) kernel texture alleles(FIG. 21). The observed segregation ratio was 5 soft:12 intermediate:3hard. In this trial, soft durum lines were especially soft with thelowest value equal to −4.

BC1 seeds were selected from six F2 plants with hardness index from 21to 49 and standard deviation greater than 28, indicating that they wereheterozygous for the Hardness containing-translocation. Seven of theseBC1F1 plants were again back-crossed blind to Havasu for BC2. As before,the ‘remnant’ F2 seeds from the self pollinated spikes were subjected toSKCS kernel texture analysis on a BC1F1 plant basis (FIG. 22). Twoplants did not fit the expected model, that is, one was soft (hardnessindex 9±12) and one was hard (hardness index 76±10). The other five wereadvanced.

Seven BC2F1 plants were grown, emasculated and back-crossed blind toHavasu, as before, for BC3. The remnant BC2F2 kernels were assayed forkernel texture (FIG. 23). Four of the seven showed a broaderdistribution (4-class histogram), whereas the other three appeared to beuniformly hard. BC3F1 seeds were selected from the softest plant(identified in FIG. 23) and advanced.

Sixteen BC3F1 plants were grown to maturity, harvested and subjected tokernel texture analysis (FIG. 24). The results showed a clear bimodaldistribution of BC3F1 plants wherein one group was comprised of BC3F1plants carrying no Hardness-containing translocation, and the otherderived from ‘soft’ and ‘hard’ kernel heterozygotes (resulting fromsegregation within the spike). The observed frequency was 6heterozygous: 10 hard.

In yet a further illustration, the following example illustrates the useof the non-transgenic Soft Svevo durum wheat WAS 080240001 as a parentto produce a non-transgenic soft textured durum wheat grain cultivar.

Plants of Soft Svevo durum wheat WAS 080240001 were grown as pollendonor for crossing to experimental durum wheat breeding line CA801-721(female). F1 seed was harvested and 28 F1 plants were grown to maturity,harvested and the seed subjected to SKCS kernel texture analysis (FIG.25).

The softest plant was selected (see FIG. 25), F2 seed was used toproduce F2 plants, four of which were harvested and subjected to SKCSkernel texture analysis (FIG. 26). The three softest plants with SKCSkernel texture less than 10 were selected and used to produce F3 plants,the seed from which were subjected to SKCS kernel texture analysis (FIG.27). The result indicated clearly that the soft kernel trait was stableand inherited from the F2 to F3.

In yet another embodiment, the following example further illustrates useof the non-transgenic Soft Svevo durum wheat WAS 080240001 was used as aparent to produce a non-transgenic soft textured durum wheat graincultivar.

Plants of Soft Svevo durum wheat WAS 080240001 were grown as pollendonor for crossing to durum wheat cultivar Alzada (female). F1 seed washarvested and 20 F1 plants were grown to maturity, harvested and theseed subjected to SKCS kernel texture analysis (FIG. 28).

Three plants were selected (see FIG. 28), F2 seed was used to produce F2plants some of which were emasculated and back-crossed blind to Alzadafor BC1. The F3 kernels from each of 26 plants were harvested andsubjected to SKCS kernel texture analysis (FIG. 29). BC1 seed from twoplants (see FIG. 29) were selected; a number of plants were grown, butonly two were subjected to SKCS kernel texture analysis (BC1F2 kernels)(FIG. 30), which indicated that both were likely segregating within thespike for Hardness. Both plants had been back-crossed blind to Alzadafor BC2.

The BC2 kernels were harvested and grown as BC2F1 plants. These plantswere emasculated and back-crossed blind to Alzada for BC3. Seven of theBC2F1 plants were subjected to SKCS kernel texture analysis (BC2F2kernels) (FIG. 31). BC3F1 kernels from the four plants with SKCShardness indexes less than 45 were selected for advancing.

In yet another embodiment, the following example illustrates further theuse of the non-transgenic Soft Svevo durum wheat WAS 080240001 as aparent to produce a non-transgenic soft textured durum wheat graincultivar.

Plants of Soft Svevo durum wheat WAS 080240001 were grown as pollendonor for crossing to the durum wheat cultivar Strongfield (female). F1seed was harvested and 18 F1 plants were grown to maturity, harvestedand the seed subjected to SKCS kernel texture analysis (FIG. 32).

F2 seeds of two F1 plants were selected and grown, and used toback-cross blind to Strongfield. BC1F1 seeds were harvested and used togrow nine BC1F1 plants. The BC1F1 plants were harvested and their BC1F2seeds subjected to SKCS kernel texture analysis (FIG. 33).

Example 5

The following example illustrates that milling of soft textured grainfrom the non-transgenic soft kernel durum wheat line.

Soft textured grain produced by a non-transgenic tetraploid wheat plantas disclosed herein (Soft Svevo), was milled on a Miag Multomatpilot-scale flour mill using only the first break rolls which were 10inches diameter, 4 inches long, with a ‘sharp’ to ‘dull’ corrugationorientation, operating at 325 rpm with a 1:2.4 roll differential, an 8%spiral with 13.5 corrugations per inch, set to a gap of 70 mm, the grainwas tempered to 14.5% moisture content for 24 hr, with an additional0.5% temper added 30 minutes prior to milling, and milled at a feed rateof 786 grams per minute. Ground products were automatically sifted ontwo 720 micron and four 145 micron sieves that are part of the Miag milland are operating at 240 rpm. The ‘overs’ from the 145 micron sieves wasmanually collected and resifted on a 150 micron sieve. Each of thevarious granulations was assayed for ash, protein and moisture contents.

With no additional work, effort, purifying or other means to increaseyield or remove bran (thereby reducing ash content), the yield ofgranular product greater than 150 microns but less than 720 microns was36.3% based on wheat to the mill, with an ash content of not more than0.74%. As CFR §137.320 provides for up to 3% of semolina may passthrough a 150 micron sieve, the total yield of semolina would equal notless than 37.3%. In contrast, Posner and Hibbs (1997, supra) indicate abreak release for durum wheat as 9% and of hard wheat as 30%, whereas inthe present embodiment, the break release for the soft durum was inexcess of 55%.

In another exemplary embodiment, soft textured grain produced by anon-transgenic tetraploid wheat plant as disclosed herein (Soft Svevo),was milled on a Miag Multomat pilot-scale flour mill employing thecomplete mill flow, the mill rolls all being 10 inches diameter, 4inches long and operating at 325 rpm with a 1:2.4 roll differential; thefirst break rolls had a ‘sharp’ to ‘dull’ corrugation orientation, an 8%spiral with 13.5 corrugations per inch, set to a gap of 70 mm; thesecond break rolls had a ‘sharp’ to ‘dull’ corrugation orientation, a10% spiral with 19 corrugations per inch, set to a gap of 12 mm; thethird break rolls had a ‘sharp’ to ‘dull’ corrugation orientation, a 10%spiral with 23.8 corrugations per inch, set to a gap of 3 mm; the first,second, third, fourth, and fifth midds rolls (reduction rolls) weresmooth (no corrugations) and were set to ‘contact’ (no gap). The grainwas tempered to 14.5% moisture content and held for 24 hr, with anadditional 0.5% temper added 30 minutes prior to milling, and milled ata feed rate of 786 grams per minute. The movement of ground products wasautomatically and pneumatically conveyed throughout the mill; productswere sifted and segregated as shown in FIG. 1. The sifter operated at240 rpm. The results of that milling are presented in Table 1. Themilling produced a yield of durum flour in excess of 80.9% at not morethan 0.82% ash.

TABLE 1 Cumulative Stream Yield % Ash % Yield % Ash % First Midds 10.30.48 10.3 0.48 First Midds Redust 3.4 0.53 13.7 0.49 First Break 10.00.55 23.7 0.51 Second Midds 25.7 0.55 49.3 0.53 Second Break 8.3 0.5957.6 0.55 Grader 3.6 0.60 61.2 0.55 Third Midds 6.4 0.91 67.6 0.58 ThirdBreak 3.5 0.92 71.2 0.59 Fourth Midds 2.7 1.74 73.9 0.64 Fifth Midds 1.03.51 74.9 0.67 Red Dog 1.5 3.78 76.4 0.73 Break Shorts 4.2 4.47 80.60.93 Reduction Shorts 0.3 5.05 80.9 0.95 Bran 19.1 5.95 100.0 1.91

A ‘straight grade’ flour comprised of the first, second and third breakstreams, the first, second, third, fourth and fifth midds streams, thefirst midds redust and grader streams, and possessed a cumulative yieldof 74.9% and a cumulative ash content of 0.67% on a dry weight basiswith a starch damage level of 3.28%, and by comparison, a commercialpastry flour had a starch damage level of 4.07%, and a commercial durumsemolina of 4.35%. The granulation of the soft durum flour being quitesimilar to that of the pastry flour (median particle size of 57 micronsversus 47 microns, soft durum flour and pastry flour, respectively), andbeing markedly finer granulation than the durum semolina (medianparticle size of 400 microns).

In another exemplary embodiment, soft textured grain produced by anon-transgenic tetraploid wheat plant as disclosed herein (Soft Svevo),was milled on a modified Brabender Quadrumat Senior mill, wherein thefirst grinding head is a Quadrumat Junior head equipped with 2.8 inchdiameter rolls possessing 18, 23, 25 and 30 corrugations, the secondgrinding head is the reduction head from a Quadrumat Senior equippedwith 2.8 inch diameter rolls possessing 36, 36, and 41 corrugations, thelast roll being smooth, both heads have been removed and areindependent. The rolls operate on a 2.14 differential at 1200 and 560rpm. The ground product from each head is collected and sifted on GreatWestern sifters using 12 inch diameter sieves, the first being clothedwith U.S. No. 32 and 100 sieves, and sifted for 2 minutes, thusproducing ‘bran’ (overs of No. 32), ‘middlings’ (the throughs of the No.32 and overs of the No. 100), and break flour (the throughs of the No.100); the middlings are then re-ground using the second head, the groundproduct of that grinding being sifted on the second sifter being clothedwith a U.S. No. 100 sieve, for 3 minutes, the throughs are ‘reductionflour’ and overs are ‘reduction shorts’. Break flour and reduction flourare combined to form ‘straight grade’ flour. The beginning grain weightis approximately 500 grams per milling. In common practice, soft wheatgrain samples are tempered to 13% moisture content, and hard wheat grainsamples to 14.5% moisture content. Durum wheats are commonly tempered to16% (Posner and Hibbs, 1997, supra).

In this exemplary embodiment, three samples of Soft Svevo grain weretempered to 13.0, 14.5 and 16.0% moisture contents for 24 hr, and milledon the aforementioned modified Quadrumat Senior mill. The result of thatmilling was as follows: at 13% temper, 46.4% break flour yield, 21.8%bran yield and 70.7% straight grade flour yield with a 0.66% ashcontent; at 14.5% temper, 41.9% break flour yield, 27.2% bran yield and66.2% straight grade flour yield with a 0.57% ash content; and at 16%temper, 38.2% break flour yield, 32.1% bran yield and 61.8% straightgrade flour yield with a 0.55% ash content. These results beingconsistent with the endosperm and general kernel characteristics beingas those of soft wheat, with an optimum temper level between 13 and14.5%.

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims.

What is claimed is:
 1. A non-transgenic tetraploid wheat plant having grain with soft textured endosperm, wherein the soft textured endosperm is expressed from puroindoline genes carried on a genome segment transferred in a 5DS(5BS) homoeologous translocation event, and wherein the non-transgenic tetraploid wheat plant having grain with soft textured endosperm has functional Ph1 and diploid like behavior in meiosis.
 2. The non-transgenic tetraploid wheat plant of claim 1, wherein the wheat plant is a durum wheat plant.
 3. The non-transgenic tetraploid wheat plant of claim 1, wherein the non-transgenic tetraploid wheat plant stably transmits the soft textured endosperm to progeny plants in normal mendelian ratios and is thereby capable of serving as a parent for breeding stable tetraploid wheat cultivars.
 4. A seed of the non-transgenic tetraploid wheat plant of claim 1, wherein the seed produces a non-transgenic tetraploid wheat plant having grain with soft textured endosperm.
 5. A non-transgenic tetraploid wheat plant, or a part thereof, produced by growing the seed of claim
 4. 6. A tissue culture of regenerable cells produced from the non-transgenic tetraploid wheat plant of claim
 1. 7. A protoplast produced from the tissue culture of claim
 6. 8. A hybrid wheat plant, having soft textured endosperm, wherein the lineage of at least one parent plant comprises a non-transgenic tetraploid wheat plant having grain with soft textured endosperm, wherein the at least one parent plant has soft textured endosperm that is expressed from puroindoline genes carried on a genome segment transferred in a 5DS(5BS) homoeologous translocation event, and wherein the non-transgenic tetraploid wheat plant having grain with soft textured endosperm has functional Ph1 and diploid like behavior in meiosis.
 9. The hybrid wheat plant of claim 8, wherein the at least one parent plant is a non-transgenic tetraploid wheat plant having grain with soft textured endosperm, wherein the at least one parent plant has soft textured endosperm that is expressed from puroindoline genes carried on a genome segment transferred in a 5DS(5BS) homoeologous translocation event, and wherein the non-transgenic tetraploid wheat plant having grain with soft textured endosperm has functional Ph1 and diploid like behavior in meiosis.
 10. A flour milled from soft textured endosperm from a non-transgenic tetraploid wheat plant having grain with soft textured endosperm, wherein the non-transgenic tetraploid wheat plant having grain with soft textured endosperm has functional Ph1 and diploid like behavior in meosis.
 11. The flour of claim 10, wherein the flour has a low level of damaged starch.
 12. The flour of claim 10, wherein the flour has a granulation similar to a pastry flour granulation.
 13. The flour of claim 12, wherein the flour has a median particle size of about 57 microns.
 14. The flour of claim 10, wherein the flour is a straight grade flour.
 15. A semolina milled from soft textured endosperm from a non-transgenic tetraploid wheat plant having grain with soft textured endosperm, wherein the non-transgenic tetraploid wheat plant having grain with soft textured endosperm has functional Ph1 and diploid like behavior in meosis.
 16. The semolina of claim 15, wherein the semolina is used for making pasta.
 17. Wheat grain kernels with soft textured endosperm, from a non-transgenic tetraploid wheat plant having grain with soft textured endosperm, wherein the non-transgenic tetraploid wheat plant having grain with soft textured endosperm has functional Ph1 and diploid like behavior in meiosis.
 18. The wheat grain kernels of claim 17, wherein the wheat grain kernels have a hardness index of between about 10 to about 45 as measured by the Perten SKCS
 4100. 